3-dimensional cellular assay platforms are increasingly recognized as robust surrogates for mimicking in vivo disease pathology. In particular, the multicellular spheroid model has been widely utilized in exploratory drug discovery campaigns. However, these complex 3D cell models have previously been restricted to low- or medium-throughput formats due to the technical logistics of forming spheroids in a 1536-well microtiter plate. We have developed a novel microphysiological 3D assay that quantitates T-cell-mediated killing of 3D colorectal cancer tumor spheroids using a new 1536-well spheroid plate. This assay incorporates CD3-stimulated primary patient T-cells in culture with colorectal cancer tumor spheroids and enables parallel assessment of spheroid size and viability as well as T-cell penetration into the 3D spheroid structure. Using this assay platform we screened a library of annotated compounds for spheroid viability and discovered several small molecule candidates that synergize with CD3 stimulation and enhance T-cell-mediated tumor spheroid killing. This phenotypic 3D cell model represents a robust organotypic ultra-HTS platform that can greatly enhance immuno-oncology drug discovery programs.

Shane Horman

Research Investigator IIIGNF

Dr. Shane Horman runs the Advanced Assay group at the Genomics Institute of the Novartis Research Foundation (GNF) in San Diego, California. He received his Ph.D. from King’s College-London in molecular genetics and was a postdoc at the University of Pennsylvania-School of Medicine and then at Cincinnati Children’s Hospital Medical Center investigating mouse models of human leukemias. Dr. Horman’s Advanced Assay group at GNF is dedicated to the development and implementation of complex and 3D high content screening platforms that may better reflect in vivo patient biology for early stage drug discovery. Dr. Horman has published numerous papers on high content 3D screening platforms and regularly presents at phenotypic drug discovery conferences.